Reporting Form from miR-1202 acts as anti-oncomiR in myeloid leukaemia by down-modulating GATA-1_S expression

Transient abnormal myelopoiesis (TAM) is a Down syndrome-related pre-leukemic condition characterized by somatic mutations in the hematopoietic transcription factor GATA-1 that result in exclusive production of its shorter isoform (GATA-1S). Given the common hallmark of altered miRNA expression profiles in haematological malignancies and the pro-leukemic role of GATA-1S, we aimed to search for miRNAs potentially able to modulate the expression of GATA-1 isoforms. Starting from an in silico prediction of miRNA binding sites in the GATA-1 transcript, miR-1202 came into our sight as potential regulator of GATA-1 expression. Expression studies in K562 cells revealed that miR-1202 directly targets GATA-1, negatively regulates its expression, impairs GATA-1S production, reduces cell proliferation, and increases apoptosis sensitivity. Furthermore, data from TAM and myeloid leukaemia patients provided substantial support to our study by showing that miR-1202 down-modulation is accompanied by increased GATA-1 levels, with more marked effects on GATA-1S. These findings indicate that miR-1202 acts as an anti-oncomiR in myeloid cells and may impact leukemogenesis at least in part by down-modulating GATA-1S levels

Supplemental Experimental Procedures from miR-1202 acts as anti-oncomiR in myeloid leukaemia by down-modulating GATA-1_S expression

Northern blot analysis and MicroRNAs cloning and capture procedur

Supplementary Tables S1-S4 from miR-1202 acts as anti-oncomiR in myeloid leukaemia by down-modulating GATA-1_S expression

Table S1. Clinical and molecular characterization of TMD/AML-DS patients with GATA-1 mutations. Table S2. Mutant sequences within or outside the putative hsa-miR-1202 binding site generated in the psiCHECK™-2 Vector GATA1 exon2 construct. Table S3. Primer sequences used for quantitative Real-time PCR analysis.Table S4. Antibodies used for western blot analysis

Altered expression of mitochondrial and extracellular matrix genes in the heart of human fetuses with chromosome 21 trisomy-2

<p><b>Copyright information:</b></p><p>Taken from "Altered expression of mitochondrial and extracellular matrix genes in the heart of human fetuses with chromosome 21 trisomy"</p><p>http://www.biomedcentral.com/1471-2164/8/268</p><p>BMC Genomics 2007;8():268-268.</p><p>Published online 7 Aug 2007</p><p>PMCID:PMC1964766.</p><p></p>ted pathways are: Oxidative Phosphorylation (cluster 1), containing 16 genes downregulated in trisomic samples, and Focal Adhesion (cluster 2), containing at least 7 genes upregulated in trisomic samples. Cluster 3 is a network of Cell Adhesion genes, mostly upregulated in trisomic samples. Downregulated genes in cluster 1 are all mitochondrial genes; upregulated genes in clusters 2 and 3 are mostly ECM genes. Green indicates downregulated genes (darker green = more downregulated); red indicates upregulated genes (darker red = more upregulated)

Altered expression of mitochondrial and extracellular matrix genes in the heart of human fetuses with chromosome 21 trisomy-3

<p><b>Copyright information:</b></p><p>Taken from "Altered expression of mitochondrial and extracellular matrix genes in the heart of human fetuses with chromosome 21 trisomy"</p><p>http://www.biomedcentral.com/1471-2164/8/268</p><p>BMC Genomics 2007;8():268-268.</p><p>Published online 7 Aug 2007</p><p>PMCID:PMC1964766.</p><p></p>H4, CDH5, CDH6 and DH1, DH3, DH4, DH5, DH6) and individual normal hearts (H1, H2, H3, H4, H5) show an inverse correlation (r = -066) between the two genes

Original blot images from miR-1202 acts as anti-oncomiR in myeloid leukaemia by down-modulating GATA-1_S expression

Transient abnormal myelopoiesis (TAM) is a Down syndrome-related pre-leukemic condition characterized by somatic mutations in the hematopoietic transcription factor GATA-1 that result in exclusive production of its shorter isoform (GATA-1S). Given the common hallmark of altered miRNA expression profiles in haematological malignancies and the pro-leukemic role of GATA-1S, we aimed to search for miRNAs potentially able to modulate the expression of GATA-1 isoforms. Starting from an in silico prediction of miRNA binding sites in the GATA-1 transcript, miR-1202 came into our sight as potential regulator of GATA-1 expression. Expression studies in K562 cells revealed that miR-1202 directly targets GATA-1, negatively regulates its expression, impairs GATA-1S production, reduces cell proliferation, and increases apoptosis sensitivity. Furthermore, data from TAM and myeloid leukaemia patients provided substantial support to our study by showing that miR-1202 down-modulation is accompanied by increased GATA-1 levels, with more marked effects on GATA-1S. These findings indicate that miR-1202 acts as an anti-oncomiR in myeloid cells and may impact leukemogenesis at least in part by down-modulating GATA-1S levels

Altered expression of mitochondrial and extracellular matrix genes in the heart of human fetuses with chromosome 21 trisomy-1

<p><b>Copyright information:</b></p><p>Taken from "Altered expression of mitochondrial and extracellular matrix genes in the heart of human fetuses with chromosome 21 trisomy"</p><p>http://www.biomedcentral.com/1471-2164/8/268</p><p>BMC Genomics 2007;8():268-268.</p><p>Published online 7 Aug 2007</p><p>PMCID:PMC1964766.</p><p></p>rom the t-test is represented on the y-axis. Genes upregulated in the trisomic samples are on the right of the horizontal axis 0 value; genes downregulated are on the left. Red dots indicate 473 genes that are significantly up- or down-regulated in the trisomic samples compared to the control samples (p < 0.05). Yellow dots indicate genes with no significant variation

Evidence of <em>Bacteroides fragilis</em> Protection from <em>Bartonella henselae</em>-Induced Damage

<div><p><em>Bartonella henselae</em> is able to internalize endothelial progenitor cells (EPCs), which are resistant to the infection of other common pathogens. <em>Bacteroides fragilis</em> is a gram-negative anaerobe belonging to the gut microflora. It protects from experimental colitis induced by <em>Helicobacter hepaticus</em> through the polysaccharide A (PSA). The aim of our study was to establish: 1) whether <em>B. fragilis</em> colonization could protect from <em>B. henselae</em> infection; if this event may have beneficial effects on EPCs, vascular system and tissues. Our <em>in vitro</em> results establish for the first time that <em>B. fragilis</em> can internalize EPCs and competes with <em>B. henselae</em> during coinfection. We observed a marked activation of the inflammatory response by Real-time PCR and ELISA in coinfected cells compared to <em>B. henselae-</em>infected cells (63 <em>vs</em> 23 up-regulated genes), and after EPCs infection with mutant <em>B. fragilis</em> ΔPSA (≅90% up-regulated genes) compared to <em>B. fragilis</em>. Interestingly, in a mouse model of coinfection, morphological and ultrastructural analyses by hematoxylin-eosin staining and electron microscopy on murine tissues revealed that damages induced by <em>B. henselae</em> can be prevented in the coinfection with <em>B. fragilis</em> but not with its mutant <em>B. fragilis</em> ΔPSA. Moreover, immunohistochemistry analysis with anti-Bartonella showed that the number of positive cells per field decreased of at least 50% in the liver (20±4 <em>vs</em> 50±8), aorta (5±1 <em>vs</em> 10±2) and spleen (25±3 <em>vs</em> 40±6) sections of mice coinfected compared to mice infected only with <em>B. henselae</em>. This decrease was less evident in the coinfection with ΔPSA strain (35±6 in the liver, 5±1 in the aorta and 30±5 in the spleen). Finally, <em>B. fragilis</em> colonization was also able to restore the EPC decrease observed in mice infected with <em>B. henselae</em> (0.65 <em>vs</em> 0.06 media). Thus, our data establish that <em>B. fragilis</em> colonization is able to prevent <em>B. henselae</em> damages through PSA.</p> </div

Morphological analysis of murine coinfected tissue by hematoxylin-eosin staining. A.

<p>Representative microscope images of hematoxylin-eosin staining of liver tissues from each group of mice uninfected, infected with <i>B. henselae</i>, <i>B. fragilis</i> and <i>B. fragilis</i> ΔPSA or coinfected, as detailed. Granulomatous inflammatory infiltrates are predominantly evident in the group of mice infected with <i>B. henselae</i> compared to the group infected with <i>B. henselae</i> and <i>B. fragilis</i> ΔPSA and particularly in the mice coinfected with <i>B. henselae</i> and <i>B. fragilis</i>. Uninfected controls are negative, as well as <i>B. fragilis</i> and <i>B. fragilis</i> ΔPSA groups. <b>B.</b> Representative sections stained with hematoxylin-eosin of aortas from all mice groups as described. In the group of mice infected with <i>B. henselae</i>, minimal lesions are observed in the aorta compared to liver tissues.</p

<i>B. fragilis</i> and <i>B. henselae</i> internalize EPCs. A.

<p>Confocal images of human early EPCs infected with <i>B. henselae, B. fragilis</i> and <i>B. fragilis</i> ΔPSA at 100 MOI. Cells were stained with anti-<i>Bacteroides</i> (red) or with anti-<i>Bartonella</i> (green) specific antibodies after 24 h from infection. <b>B.</b> EPCs coinfected with <i>B. henselae,</i> and <i>B. fragilis</i> or <i>B. fragilis</i> ΔPSA as indicated. A MOI of 100 was used for all bacteria strains. Cells were stained with anti-<i>Bacteroides</i> (red) and with anti-<i>Bartonella</i> (green) specific antibodies after 24 h from infection.</p